Salivary markers of systemic diseases pptPresentation Transcript
SALIVARY MARKERS OF SYSTEMIC DISEASES BY Y.PRAGZNA(10T21S0113) UNDER THE GUIDANCE OF Mrs. M. SANDHYA M.PHARM ASSOCIATE PROFESSOR
Advantages & disadvantages
Saliva collection, processing & storage
Salivary proteome & transcriptome
New technologies of measurement
BIOMARKERS: Biomarkers are defined as cellular, biochemical and molecular characteristics by which normal/abnormal processes can be recognized and/or monitored.
SALIVARY MARKERS Non invasive diagnosis of disease and monitoring of general health
Saliva is the mixed product of 3 major salivary glands.
Parotid Sub mandibular Sub lingual
It is slightly acidic(pH 6-7).
Several physiological & pathological conditions modify saliva production quantitatively.
Functions of saliva PROTECTIVE
FOOD & SPEECH RELATED
Prevention of caries
SALIVA AS A DIAGNOSTIC FLUID ADVANTAGES
It is safe, non-invasive and simple.
No needles and No cringing.
It has an average protein concentration of 1.5-2 mg/ml.
Cost of shipping & Storage tend to be lower.
Informative analytes are present in lower amounts in saliva than in serum. DISADVANTAGES
Whole saliva collection Ductal secretions collection Parotid secretions collection Sub mandibular & sub lingual secretions collection Saliva collection
PROCESSING & STORAGE: All procedures must be performed on ice: (1)Divide the saliva samples into multiple 330μL samples placed in cryotubes at -80ºc. Further processed for
Protein analysis: Add protease inhibitors like aprotinin(0.33µL), 3.3µL PMSF.
RNA analysis: Add 1.65µL of SUPERase inhibitor.
(2)Invert gently to mix. (3)Store all fractions at -80ºc.
Salivary/Oral Fluid Biomarkers Possibilities of Use DNA Standard genotyping Genetic information of the hosting human body Bacterial infection (oral microbes in the mouth) Diagnosing carcinomas of head & neck Forensics mRNA Information on transcription rates RNA Viral/bacterial identification Proteins Genetic information Translational regulation Diagnosing periodontitis Detecting dental cavities Immunoglobulin Diagnosing viruses Metabolites Diagnosing periodontitis
DIAGNOSTIC APPLICATIONS OF SALIVA:
Intra cellular pathways
Active & Passive diffusion
Extra cellular pathways
Membrane of acinar
& ductal cells
There are raised concentrations of NaCl, IgA, IgG, lactoferrin, inflammatory mediators such as PGE2, TXB2, Interleukin-6 etc.
Saliva of CF patients contain increased calcium & phosphate levels, urea & uric acid and abnormally elevated levels of PGE2.
It effects flow rate & composition of saliva. Salivary alterations includes higher levels of IgA, Salivary Peroxidase, glucose content, K, Salivary total protein & amylase.
ORAL MUCOSAL DISEASES:
Nitric oxide levels were increased.
P53 antibody can be detected in oral squamous cell carcinoma, increased levels of salivary kallikrein levels in oral malignant tumors, CA 15-3 in breast cancer. CVS: Determination of salivary amylase activity before 6hrs after cv surgery. KIDNEY DYSFUNCTION: Hypo salivation , changes in taste, ammonium smelling breath, oral mucosal pain. EPILEPSY: Gingival hypertrophy, IgA deficiency
VIRAL DISEASES:Acute hepatitis A & B were diagnosed based on the presence of IgM antibodies in saliva.HIV:Salivary IgA levels.BACTERIAL INFECTIONS:By monitoring the presence of antibodies to microorganisms found in saliva & oral cavity
AGAIN & AGAIN ABOUT TOBACCO:
Smoking causes decrease in salivary important antioxidant levels.
Salivary nicotine levels are considered to be the most reliable marker
Drug monitoring ANTI-ASTHMATICS:Theophylline ANTI EPILEPTICS: Carbamazapine, Diazepam, Ethosuximide, Lamotrigine, Topiramate, Phenytoin, Primidone, Valproic acid ANTI ARRYTHMIC/ANTI HYPERTENSIVES: Digoxin, Metoprolol, and Procainamide. ANTI MICROBIALS:Clarithromycin, Gentamycin, Isoniazid, Ofloxacin, Sulfanilamide ANTIPYRETICS: Acetaminophen, Antipyrine, Paracetamol ANTI NEOPLASTICS:Cisplatin, Methotrexate, Texol, Topotecan ANTI VIRAL: Quinine LITHIUM PSYCHOTROPIC:Lithium, Amitriptyline MISCELLANEOUS: Nicotine, Ethanol, Caffeine, Tolbutamide, Quinine ILLICIT DRUGS: Amphetamine, Barbiturates, Benzodiazepines, Opiods, Cocaine, Phencyclidine
HORMONE MONITORING: Steroids like cortisol, aldosterone, testosterone, insulin SIALOCHEMISTRY ANALYSIS: Early monitoring of an exposure to lead, cadmium, mercury because of higher salivary levels of these elements
CLASSIFICATION OF SALIVARY BIO MARKERS:
Diagnostic molecular targets SALIVARY PROTEOME: Use of ‘Divide & conquer’ strategy
Initial fractionation of proteins with separation techniques
Digestion with proteolytic enzyme, trypsin
Analysis by 1-D, 2-D LC-MS
Finally data is processed & submitted for database searching
SALIVARY PROTEOMIC ANALYSIS
It includes mRNA molecules that cells use to convey the instructions carried by DNA for subsequent protein production.
At present, the main strategy for identification of salivary transcriptomic biomarkers is through microarray technology.
SALIVARY TRANSCRIPTOME: DNA MICROARRAY TECHNOLOGY
NEW TECHNOLOGIES FOR MEASURING SALIVARY BIOMARKERS:
In this POC device, saliva(100-300μL) is placed into the salivary collection module, and then delivered into the Nano-biochip.
Micro pits within a silicon wafer are populated with a variety of chemically sensitized bead micro reactors.
MICRO BEAD ARRAY TECHNIQUE NANO-BIOCHIP
CONCLUSION: The components of saliva act as a “Mirror of the body’s health”, widespread use and growing acceptability of saliva as a diagnostic tool is helping individuals, researchers, health care professionals and community health programs to better detect and monitor diseases and to improve the general health of the public
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